The Biological Research Of Astrocytes From Primary Culture And Neural Stem Cells Induced

Objective:Astrocyte is the largest number of glial cells in the central nervous system and plays an important role in many functions. There are two methods to get astrocytes in vitro, primary culture and neural stem cells induced. But the biological characteristics of astrocytes from two methods need to be further studied. In this study, through cell growth, the reaction of scratch and detection cell function and carcinogenicity, etc., directly to verify whether the biology of astrocytes from primary cultured and neural stem cells induced is equivalent.Methods:Through primary culture we induced neural stem cells from neonatal rat hippocampal into astrocytes, and separate the cerebral cortex to get astrocytes with classical method. The two kinds of astrocytes were purified with differential adherent and thermostatic shaker oscillation, and the purity was detected by indirect immunofluorescence. Two types of cells in cell growth and the response of scratch were compared. And using fluorescence quantitative PCR and Western-blot, we detect intracellular protein molecules for the comparison of the two biological properties. Apply SPSS17.0statistical software to analyze, all measurement data were expressed as mean standard deviation (X±s), two groups use independent samples t-test, P<0.05was considered statistically significant.Result:The methods of primary cultured and neural stem cells induced both can get astrocytes. Confirmed by indirect immunofluorescence experiments, the purity of astrocytes from neural stem cell induced can reach99.4±0.5%, while the cell purity from traditional methods is94.2±2%. There are no significant differences of astrocytes from the two methods in morphology, growth trends and response to injury. Using Western-blot analysis of the expression intracellular EGFR and P53protein and quantitative PCR determination of GFAP expression, there are no significant differences in the expression levels of the two types of cells either.Conclusion:Through culture and purification astrocytes induced from neural stem cells can be pure astrocytes. There is no significant difference of astrocytes by the two methods in cell growth, the response of scratch, functional identification and carcinogenicity, and the way of neural stem cells induced differentiation has the characteristics of easy preparation and purification. Therefore it can be used as the conventional method of preparation of astrocytes for every kind of astrocytes tests in vitro and in vivo.